A high-yield isolation and enrichment strategy for human lung microvascular endothelial cells.

Abstract:

:Vasculopathies, characterized by the formation of fragile and abnormal microvessels, are associated with the severity of many chronic lung diseases, including pulmonary fibrosis, emphysema/chronic obstructive pulmonary disease, systemic sclerosis, and hypertension. However, the study of human lung vasculature has been limited by the ability to isolate generous quantities of microvascular endothelial cells (MVEC) free from mesenchymal contamination. Expansion and passaging of primary human MVEC in vitro typically results in loss of a traditional phenotype in favor of an intermediate mesenchymal one, as early as passage five. Here we provide a detailed protocol for the selection of large quantities of enriched primary human lung MVEC based upon differential adherence from mesenchyme and simple magnetic separation, which decreases the need for excessive passaging, in order to obtain sufficient cell numbers to successfully freeze stock cultures. Additional protocols are provided for Ac-di-LDL selection, characterization, and a sandwich angiogenesis method of functional tube formation. The complete protocol including cell isolation and characterization takes approximately six weeks to complete.

journal_name

Pulm Circ

journal_title

Pulmonary circulation

authors

Gaskill C,Majka SM

doi

10.1177/2045893217702346

subject

Has Abstract

pub_date

2017-03-27 00:00:00

pages

108-116

issue

1

eissn

2045-8932

issn

2045-8940

pii

10.1177_2045893217702346

journal_volume

7

pub_type

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